Non-viral gene therapy vectors are particularly attractive for a large number of reasons, yet efforts to build stable and effective ones have been largely unsuccessful to date. The ideal non-viral vector would have small size, be non-aggregating under physiologic conditions, targetable to specific cell types, highly efficient at gene delivery and easy to construct. During Phase I, a new non-viral method for constructing stable, non- aggregating DNA-containing particles was developed. These particles were formed by a process termed template polymerization in which cationic monomers were polymerized along a DNA template. The covalent attachment of hydrophilic polymers to the cationic monomers prior to template polymerization resulted in steric stabilization of the formed particles of condensed DNA. This breakthrough technology allows for the first time the formation of condensed DNA containing particles that are stable under physiological salt conditions. In phase II the basic particle formation strategy (developed during phase I) will be expanded using parallel synthetic techniques in a focused effort to create highly effective gene delivery particles. Optimized particles formed in this manner will be tested for their ability to deliver reporter genes into cells in vitro and in vivo and subsequently to deliver therapeutic genes into small animal disease models. We have obtained preliminary data on in vitro and in vivo gene delivery using these particles. PROPOSED COMMERCIAL APPLICATION: This new method of non-viral gene delivery will be commercialized by: l) selling the technology in the form of reagent kits to biomedical and basic researchers 2) developing the technology "in-house" for gene therapy of genetic, metabolic, and acquired liver diseases 3) licensing the technology for in vivo gene delivery to pharmaceutical companies.